Difference between revisions of "Part:BBa K274120:Experience"
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
how you used this part and how it worked out.
 
how you used this part and how it worked out.
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Characterization by UiOslo_Norway 2019
 
Characterization by UiOslo_Norway 2019
  
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This part was of special interest to our team. The part consists of homologs of the genes we used; crtE, crtB and crtI from Deinococcus radiodurans. BBa_K274120 has individual ribosomal binding sites for each individual enzyme. The part that we made had a single ribosomal binding site followed by all three genes in a single open reading frame. The first two genes had their stopcodons removed (see BBa_K2971010).  
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This part was of special interest to our team. The part consists of homologs of the genes we used; <i>crtE, crtB</i> and <i>crtI</i> from Deinococcus radiodurans. BBa_K274120 has individual ribosomal binding sites for each individual enzyme. The part that we made had a single ribosomal binding site followed by all three genes in a single open reading frame because the first two genes lacked stop codons (see BBa_K2971004 and BBa_K2971010). Figure 1 shows the result of expressing BBa_K274120 in <i>E.coli</i> (BL21) for 24 hours. The red coloration in the induced pellet is caused by the presence of lycopene. The induced culture with BBa_K274120 produced significant amount of lycopene when compared to expression of BBa_K2971004 (figure 2).
  
 
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<img style="width:40% !important;" src="https://2019.igem.org/wiki/images/9/99/T--UiOslo_Norway--HQ%26pBad.jpg?">
 
<img style="width:40% !important;" src="https://2019.igem.org/wiki/images/9/99/T--UiOslo_Norway--HQ%26pBad.jpg?">
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<strong>Figure 1: Induction of BBa_K274120 in Escherichia coli (BL21)</strong></br>
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Expression was induced in 2&#x25; arabinose </br>
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<strong>Figure 1: Induction of BBa_K274120 in <i>Escherichia coli</i> (BL21)</strong></br>
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From left to right &#58;Uninduced E.coli with empty expression vector, Induced E.coli with empty expression vector, uinduced E.coli with BBa_K274120, induced E.coli with BBa_K274120.
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Expression was induced with 2&#x25; arabinose </br>
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From left to right &#58;Uninduced <i>E.coli</i> with empty expression vector, induced <i>E.coli</i> with empty expression vector, uninduced <i>E.coli</i> with BBa_K274120, induced <i>E.coli</i> with BBa_K274120.
 
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<img style="width:40% !important;" src="https://2019.igem.org/wiki/images/4/4e/T--UiOslo_Norway--our%26pBad.jpg">
 
<img style="width:40% !important;" src="https://2019.igem.org/wiki/images/4/4e/T--UiOslo_Norway--our%26pBad.jpg">
 
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<strong>Figure 2: Expression of crtEBI in Escherichia coli (BL21)</strong></br>
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<strong>Figure 2: Expression of crtEBI in <i>Escherichia coli</i> (BL21)</strong></br>
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Expression was induced in 2&#x25; arabinose </br>
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Expression was induced with 2&#x25; arabinose </br>
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From left to right &#58;Uninduced E.coli with empty expression vector, Induced E.coli with empty expression vector, induced E.coli with crtEBI expression vector, uninduced E.coli with crtEBI expression vector.
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From left to right &#58;Uninduced <i>E.coli</i> with empty expression vector, induced <i>E.coli</i> with empty expression vector, induced <i>E.coli</i> with crtEBI expression vector, uninduced <i>E.coli</i> with crtEBI expression vector.
 
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Since the production of lycopene was so high in BBa_K274120 compared to our composite part we tried using the culture in a simple solar cell (figure 3). The design of the solar cell is comparable to a regular dye sensitized solar cell (see our design page).
  
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<img style="width:40% !important;" src="https://2019.igem.org/wiki/images/c/c0/T--UiOslo_Norway--Solarcell.jpg">
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<strong>Figure 3: Pelleted culture of E.coli expressing BBa_K274120 used in a simple solar cell</strong></br>
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Expression was induced in 2&#x25; arabinose. </br> We did not observe any detectable current from this solar cell.
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Latest revision as of 17:00, 21 October 2019

_NOTOC_ This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K274120

User Reviews

Characterization by UiOslo_Norway 2019

This part was of special interest to our team. The part consists of homologs of the genes we used; crtE, crtB and crtI from Deinococcus radiodurans. BBa_K274120 has individual ribosomal binding sites for each individual enzyme. The part that we made had a single ribosomal binding site followed by all three genes in a single open reading frame because the first two genes lacked stop codons (see BBa_K2971004 and BBa_K2971010). Figure 1 shows the result of expressing BBa_K274120 in E.coli (BL21) for 24 hours. The red coloration in the induced pellet is caused by the presence of lycopene. The induced culture with BBa_K274120 produced significant amount of lycopene when compared to expression of BBa_K2971004 (figure 2).

Figure 1: Induction of BBa_K274120 in Escherichia coli (BL21)
Expression was induced with 2% arabinose
From left to right :Uninduced E.coli with empty expression vector, induced E.coli with empty expression vector, uninduced E.coli with BBa_K274120, induced E.coli with BBa_K274120.

Figure 2: Expression of crtEBI in Escherichia coli (BL21)
Expression was induced with 2% arabinose
From left to right :Uninduced E.coli with empty expression vector, induced E.coli with empty expression vector, induced E.coli with crtEBI expression vector, uninduced E.coli with crtEBI expression vector.

Since the production of lycopene was so high in BBa_K274120 compared to our composite part we tried using the culture in a simple solar cell (figure 3). The design of the solar cell is comparable to a regular dye sensitized solar cell (see our design page).

Figure 3: Pelleted culture of E.coli expressing BBa_K274120 used in a simple solar cell
Expression was induced in 2% arabinose.
We did not observe any detectable current from this solar cell.


UNIQa3c2d40d9f2922c8-partinfo-00000002-QINU UNIQa3c2d40d9f2922c8-partinfo-00000003-QINU